Study conducted on hAPP tg mice which overexpress human APP751 with the London (V717I) and Swedish (K670M/N671L) mutations under the regulatory control of the neuron-specific murine (m)Thy-1 promoter (gift of Dr. Eliezer Masliah).
Number and surface area of amyloid plaques were evaluated in both cortex and hippocampus. 6E10 (Signet; 1:5000; monoclonal) as primary antibody and Cy3 fluorescent goat anti-mouse IgG as secondary antibody (Jackson ImmunoResearch) were used for Ab staining. Sections of the same brains were also used for synaptophysin staining (Chemicon; 1:5000; monoclonal). Ab1-40 and Ab1-42 were assayed using commercially available ELISA kits (The Genetics Company).
Antibodies used for Western blots were R1736 (specific for sAPP-a; a gift from Dr. D.J. Selkoe), Ab14 (PS1-NTF; a gift from Dr. S.E. Gandy), pen-2 (a gift from Dr. S.S. Sisodia), N-Cadherin (C32, C-terminal; BD Transduction Laboratories), ApoE (Santa Cruz Biotechnology), b-tubulin (Sigma), BACE1, and Nicastrin (Affinity BioReagents). Rbt × APP (CTF), 22C11 (APP-NTF), and Rbt × PS1 (PS1-CTF) were from Chemicon International. BACE stainings (N-terminal; Affinity BioReagents) were confirmed with two additional antibodies; Ms × BACE1 (Chemicon International) and BACE (C-terminal; Affinity BioReagents).